Tumour Necrosis Factor (TNF) is a cytokine which is produced initially as a cell-associated 28 kD precursor. It is released as an active, 17 kD form (D-M Jue et al, (1990) Biochemistry, 29:8371-8377), which can mediate a large number of deleterious effects in vivo. When administered to animals or humans it causes inflammation, fever, cardiovascular effects, haemorrhage, coagulation and acute phase responses, similar to those seen during acute infections and shock states. Chronic administration can also cause cachexia and anorexia. Accumulation of excessive TNF can be lethal.
There is considerable evidence from animal model studies that blocking the effects of TNF with specific antibodies can be beneficial in acute infections, shock states, graft versus host reactions and autoimmune disease. TNF is also an autocrine growth factor for some myelomas and lymphomas and can act to inhibit normal heamatopoiesis in patients with these tumours.
Preventing the production or action of TNF is, therefore, predicted to be a potent therapeutic strategy for many inflammatory, infectious, immunological or malignant diseases. These include, but are not restricted to, septic shock, haemodynamic shock and sepsis syndrome (Mathison et al, (1988) J. Clin. Invest. 81:1925-1937; Miethke et al (1992), J. Exp. Med. 175:91-98), post ischaemic reperfusion injury, malaria (Grau et al (1989), Immunol. Rev. 112:49-70); mycobacterial infection (Barnes et al (1992) Infect. Imm. 60:1441-6), meningitis, psoriasis, congestive heart failure, fibrotic disease, cachexia, graft rejection, cancer, autoimmune disease, rheumatoid arthritis, multiple sclerosis, radiation damage, toxicity following administration of immunosuppressive monoclonal antibodies such as OKT3 or CAMPATH-1 and hyperoxic alveolar injury.
Current clinical anti-TNF strategies involve the use of corticosteroids such as dexamethasone, and the use of cyclosporin-A or FK506, which are non-specific inhibitors of cytokine gene transcription. Phosphodiesterase inhibitors such as pentoxyfilline have been shown to be more specific inhibitors of TNF gene transcription (Endres S . (1991) Immunol. 72:56-60, Schandene et al (1992), Immunol. 76:30-34, Alegre M L, et al (1991); Transplantation 52:674-679, Bianco et al (1991) Blood 78:1205-1221). Thalidomide has also been shown to inhibit TNF production by leucocytes (Sampajo et al (1991), J. Exp. Med. 173:699-703). In experimental settings, anti-TNF monoclonal antibodies, soluble TNF receptors and soluble TNF receptor/immunoadhesins have been shown to specifically inhibit the effects of TNF action (Bagby et al (1991) J. Infect. Dis. 163:83-88, Charpentier et al. (1991) Presse-med. 20:2009-2011, Silva et al (1990) J. Infect. Dis. 162:421-427; Franks et al (1991) Infect. Immun. 59:2609-2614, Tracey et al (1987) Nature 330:662-664; Fischer et al (1992) PNAS USA in press, Lesslauer et al (1991) Eur. J. Immunol. 21:2883-2886, Ashkenazi et al (1991) PNAS USA 88:10535-10539).
It has recently been shown that the effects of TNF are mediated by two peptides, TNF.alpha. and TNF.beta.. Although these peptides have only 30% homology with each other, they activate the same receptors and are encoded by immediately adjacent genes. As used herein, the term tumour necrosis factor or TNF therefore means tumour necrosis factor a and peptides having a high degrees of sequence homology with, or substantially similar physiological effects to, TNF.alpha., for example TNF.beta..
One of the objectives of the present invention is to provide compounds which substantially inhibit the release of TNF from cells, and therefore may be used in the treatment of conditions mediated by TNF. Such uses include, but are not limited to, the treatment of inflammation, fever, cardiovascular effects, haemorrhage, coagulation and acute phase response, cachexia and anorexia, acute infections, shock states, graft versus host reactions and autoimmune disease.
In normal tissues, cellular connective tissue synthesis is offset by extracellular matrix degradation, the two opposing effects existing in dynamic equilibrium. Degradation of the matrix is brought about by the action of proteinases released from resident connective tissue cells and invading inflammatory cells, and is due, in part, to the activity of at least four groups of metalloproteinases. These are the collagenases (interstitial collagenase, MMP-1; PMN collagenase, MMP-8, collagenase-3, MMP-13), the gelatinases (gelatinase A, MMP-2, 72 kDa-gelatinase, Type IV collagenase; gelatinase B, MMP-9, 92 kDa-gelatinase, Type IV collagenase) the stromelysins (proteoglycanase, MMP-3, stromelysin-1, transin; stromelysin-2, MMP- 10; stromelysin 3, MMP- 11) and the membrane type matrix metalloproteinases (MT-1, MMP-14; MT-2, MMP-15; MT-3, MMP-16 and MT-4, MMP-17). Normally these catabolic enzymes are tightly regulated at the level of their synthesis and secretion and also at the level of their extracellular activity, the latter through the action of specific inhibitors, such as TIMP (tissue inhibitors of metalloproteinase), which form inactive complexes with metalloproteinases, and more general proteinase inhibitors such as .alpha..sub.2 -macroglobulins.
The accelerated, uncontrolled breakdown of connective tissues by metalloproteinase catalysed resorption of the extracellular matrix is a feature of many pathological conditions such as rheumatoid arthritis, osteoarthritis, septic arthritis, corneal, epidermal or gastric ulceration; tumour metastasis or invasion; periodontal disease, proteinuria, coronary thrombosis associated with atherosclerotic plaque rupture and bone disease. Inhibitors may also be useful in preventing the pathological squaelae following a traumatic injury that could lead to a permanent disability. These compounds may also have utility as a means for birth control by preventing ovulation or implantation. It can be expected that the pathogenesis of such diseases is likely to be modified in a beneficial manner by the administration of metalloproteinase inhibitors and numerous compounds have been suggested for this purpose [for a general review see R C Wahl, et al Ann. Rep, Med. Chem. 25, 175-184, Academic Press Inc., San Diego (1990)].
Compounds which have the property of inhibiting the action of metalloproteinases involved in connective tissue breakdown such as collagenase, stromelysin and gelatinase have been shown to inhibit the release of TNF both in vitro and in vivo (A J H Gearing et al (1994), Nature, 370:555-557; G M McGeehan at al (1994), Nature, 370:558-561: M J Crimmin et al, WO 93/20047 and WO 94/10990). All of these reported inhibitors contain a hydroxamic acid zinc binding group.
It is, therefore, a further objective of this invention to provide compounds which, in addition to inhibiting TNF release, also may inhibit the action of certain MMPs, and hence may be used in the treatment of patients who suffer from conditions mediated by TNF and/or MMPs such uses include, but are not limited to inflammation, fever, cardiovascular effects, haemorrhage, coagulation and acute phase response, cachexia and anorexia, acute infections, shock states, graft versus host reactions and autoimmune disease; and those involving tissue breakdown such as bone resportion, inflammatory diseases, dermatological conditions, tumour growth, angiogenesis and invasion by secondary metastases, in particular rheumatoid arthritis, osteoarthritis, periodontitis, gingivitis, corneal ulceration, tumour growth, angiogenesis and invasion by secondary metastases.
Compounds that inhibit collagenase, which possess structural portions akin to those of the instant invention include those encompassed by U.S. Pat. No. 4,511,504 issued Apr. 16, 1985; U.S. Pat. No. 4,568,666, issued Feb. 4, 1986.
Compounds of related structure that are claimed to inhibit stromelysin (proteoglycanase) are encompassed by U.S. Pat. No. 4,771,037, issued Sep. 13, 1988.
The applicants believe that stromelysin and collagenase inhibitors have utility in preventing articular cartilage damage associated with septic arthritis. Bacterial infections of the joints can elicit an inflammatory response that may then be perpetuated beyond what is needed for removal of the infective agent resulting in permanent damage to structural components. Bacterial agents have been used in animal models to elicit an arthritic response with the appearance of proteolytic activities. See J. P. Case et al (1989), J. Clin. Invest., 84:1731-40; R. J. Williams at al (1990), Arth. Rheum., 33:533-41.
The applicants also believe that inhibitors of stromelysin, collagenase and gelatinase will be useful to control tumour metastasis, optionally in combination with current chemotherapy and/or radiation. See L. M. Matrisian et al (1986), Proc. Natl. Acad. Sci., USA, 83:9413-7; S. M. Wilhelm et al (1987), Ibid. 84:6725-29; Z. Werb et al (1989), J. Cell Biol., 109:872-889; L. A. Liotta et al (1983), Lab. Invest., 49:636-649; R. Reich et al in Metatasis; Ciba Foundation Symposium, Wiley, Chicester, 1988, pp. 193-210.
Secreted proteinases such as stromelysin, collagenase and gelatinase play an important role in processes involved in the movement of cells during metastatic tumour invasion. Indeed, there is also evidence that the matrix metalloproteinases are over expressed in certain metastatic tumour cell lines. In this context, the enzyme functions to penetrate underlying basement membranes and allow the tumour cell to escape from the site of primary tumour formation and enter the circulation. After adhering to blood vessel walls, the tumour cells use these same metalloproteinases to pierce underlying basement membranes and penetrate other tissues, thereby leading to tumour metastasis. Inhibition of this process would prevent metastasis and improve the efficacy of current treatments with chemotherapeutics and/or radiation.
These inhibitors should also be useful for controlling periodontal diseases, such as gingivitis. Both collagenase and stromelysin activities have been isolated from fibroblasts derived from inflamed gingiva (V. J. Uitto et al (1981), J.Periodontal Res., 16:417-424). Enzyme levels have been correlated to the severity of gum disease; C. M. Overall et al (1987), J. Periodontal Res., 22:81-88.
Proteolytic processes have also been observed in the ulceration of the cornea following alkali bums (S. I. Brown et al (1969), Arch. Opthalmol., 81:370-373). Mercapto-containing peptides do inhibit the collagenase isolated from alkali-burned rabbit cornea (F. R. Burns et al (1989), Invest. Opthalmol, 30:1569-1575). Treatment of alkali-burned eyes or eyes exhibiting corneal ulceration as a result of infection with inhibitors of these metalloendoproteinases in combination with sodium citrate or sodium ascorbate and/or antimicrobials may be effective in preventing developing comeal degradation.
Stromelysin has been implicated in the degradation of structural components of the glomerular basement membrane (GBM) of the kidney, the major function of which is to restrict passage of plasma proteins into the urine (W. H. Baricos et al (1989), Biochem. J., 254:609-612). Proteinuria, a result of glomerular disease, is excess protein in the urine caused by increased permeability of the GBM to plasma proteins. The underlying causes of the increased GBM permeability are unknown, but proteinases including stromelysin may play an important role in glomerular diseases. Inhibition of this enzyme may alleviate the proteinura associated with kidney malfunction.
It is suggested that inhibition of matrix metalloproteinase activity may prevent the rupturing of atherosclerotic plaques leading to coronary thrombosis. The tearing or rupture of atherosclerotic plaques is the most common event initiating coronary thrombosis. Destabilisation and degradation of the connective tissue matrix surrounding these plaques by proteolytic enzymes or cytokines released by infiltrating inflammatory cells has been proposed as a cause of plaque fissuring. Such tearing of these plaques can cause an acute thrombolytic event as blood rapidly flows out of the blood vessel. High levels of stromelysin RNA message have been found to be localised to individual cells in atherosclerotic plaques removed from heart transplant patients at the time of surgery (A. M. Henney et al (1991), Proc. Nat'l. Acad. Sci. USA, 88:8154-8158). Inhibition of matrix metalloproteinases by these compounds may aid in preventing or delaying the degradation of the connective tissue matrix that stabilises the atherosclerotic plaques, thereby preventing events leading to acute coronary thrombosis.
It has been recently shown in a model of congestive heart failure (CHF) in the pig, that during CHF the are marked changes in the morphological structure of the heart. Ventricular dilation and wall thinning caused by changes to the extracellular matrix results in fewer collagen connections between cardiomyocytes and less total collagen. In such an instance a weaker force of contraction leads to an inefficient ventricular operation. It is believed that specific inhibitors of matrix metalloproteinases will play a key role in stabilising the extracellular matrix and therefore be important in the treatment and/or prevention of CHF.
It has recently been shown (WO 96/0240) that inhibitors of the matrix metalloproteinases, such as collagenase and stromelysin also inhibit the formation of human soluble CD23. CD23 is a 45 kDa type II integral protein expressed on the surface of a variety of mature cells, including B and T lymphocytes, macrophages, NK cells, Langerhans cells, monocytes, eosinophils and platelets (Delespesse et al, Adv. Immunology, 49, 1991, 149; Grangette et al, J., Immunol, 143, 1989, 3580). Several activities have been ascribed to soluble CD23 in man, all of which involve IgE regulation. Particular activities include:
i) antigen presentation PA0 ii) IgE mediated eosinophil cytotoxicity PA0 iii) B cell homing to lymph nodes and the spleen PA0 iv) downregulation of IgE synthesis PA0 R.sup.1 is a C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.1-6 alkyl- aryl, aryl, C.sub.1-6 alkyl-heteroaryl, heteroaryl or C.sub.1-6 alkyl-AR.sup.9 group where A is O, NR.sup.9 or S(O).sub.m where m=0-2, and R.sup.9 is H, PA0 C.sub.1-4 alkyl, aryl, heteroaryl, C.sub.1-4 alkyl- aryl or C.sub.1-4 alkyl-heteroaryl. If A.dbd.NR.sup.9 the PA0 R.sup.9 groups may be the same or different. PA0 R.sup.2 is hydrogen or a C.sub.1-6 alkyl group; PA0 R.sup.3 is a [Alk].sub.n R.sup.6 group where Alk is a C.sub.1-6 alkyl or C.sub.2-6 alkenyl group and n is zero or an integer 1; PA0 X is a group NR.sup.4 R.sup.5 where R.sup.4 is hydrogen or an aryl, heteroaryl, C.sub.1-6 alkyl-heteroaryl, cyclo(C.sub.3-6)alkyl, C.sub.1-6 alkyl-cyclo(C.sub.3-6)alkyl, heterocyclo(C.sub.4-6)alkyl (such as pyrrolidine or piperidine) or C.sub.1-6 alkyl-heterocyclo(C.sub.4-6)alkyl group or the group C.sub.1-6 alkyl optionally substituted by amino (NH.sub.2), aryl, arylamino, protected amino, di(C.sub.1-6 alkyl)amino, mono(C.sub.1-6 alkyl)amino, CO.sub.2 H, protected carboxyl, carbamoyl, mono(C.sub.1-6 alkyl) carbamoyl, di(C.sub.1-6 alkyl) carbamoyl, and R.sup.5 is hydrogen; NR.sup.4 R.sup.5 may also form a ring such as pyrrolidino, piperidino or morpholino group. PA0 R.sup.7 is aryl (optionally substituted with R.sup.10), heteroaryl (optionally substituted with R.sup.10), C.sub.1-5 alkyl (optionally substituted with R.sup.10), C.sub.1-5 alkyl-aryl (optionally substituted with R.sup.10), C.sub.1-5 alkyl-heteroaryl (optionally substituted with R.sup.10), cyclo (C.sub.3-6) alkyl (optionally substituted with R.sup.10), cyclo (C.sub.3-6)alkenyl (optionally substituted with R.sup.10), C.sub.1-5 alkyl-cyclo (C.sub.3-6) alkyl (optionally substituted with R.sup.10), the groups ##STR2## where p=1-2, or the group ##STR3## where B and C may be selected from the groups O,S,C(R.sup.9).sub.2, or NR.sup.9 and these may be the same or different; PA0 When Y is C.dbd.O, R7 can also be CO.sub.2 R.sup.2 ; PA0 R.sup.8 is hydrogen; PA0 R.sup.6 is an optionally substituted cyclo (C.sub.3-6) alkyl, cyclo (C.sub.3-6) alkenyl, C.sub.1-6 alkyl, aryl, C.sub.1-6 alkoxy-aryl, benzyloxyaryl, heteroaryl, COR.sup.9, C.sub.1-3 alkyl-aryl, C.sub.1-3 alkyl-heteroaryl, C.sub.1-6 alkyl-CO.sub.2 R.sup.9, C.sub.1-6 alkyl-NHR.sup.9, AR.sup.9, CO.sub.2 R.sup.2, CONHR.sup.12, NHCOR.sup.12, NHCO.sub.2 R.sup.12, NHSO.sub.2 R.sup.12, amidine or guanidine group; PA0 where R.sup.12 is a group C.sub.1-4 alkyl, C.sub.1-4 alkylaryl, C.sub.1-4 alkyl-heteroaryl, cyclo (C.sub.3-6) alkyl, C.sub.1-4 alkyl-cyclo (C.sub.3-6) alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkenyl-aryl, aryl or heteroaryl as defined above. PA0 R.sup.10 is AR.sup.9, COR.sup.13, SO.sub.2 R.sup.9 (where R.sup.9 is not H), SO.sub.2 N(R.sup.9).sub.2, NR.sup.9 R.sup.11, COR.sup.9, phthalimido or succinimido; PA0 R.sup.11 is a COR.sup.9, CO.sub.2 R.sup.2 (where R.sup.2 is not H), CONHR.sup.9 or SO.sub.2 R.sup.9 (where R.sup.9 is not H) group; PA0 R.sup.13 is a OH, OC.sub.1-4 alkyl, O(C.sub.1-4 alkyl) aryl, N(R.sup.9).sub.2 (in which R.sup.9 is the same or different); PA0 Y is C.dbd.O, C.dbd.S or SO.sub.2 ; PA0 R.sup.1 is C.sub.1-6 alkyl or C.sub.1-4 alkyl-AR.sup.9 where A is S(O).sub.m, NR.sup.9, or O and m=0,1 or 2, and R.sup.9 is H, C.sub.1-4 alkyl heteroaryl or aryl; PA0 R.sup.2 is H or C.sub.1-4 alkyl; PA0 R.sup.3 is [Alk].sub.n R.sup.6 where n=0 or 1, Alk is C.sub.1-4 alkyl and R.sup.6 is C.sub.1-4 alkyl, C.sub.1-3 alkyl-aryl C.sub.1-3 alkyl-heteroaryl, AR.sup.9, CO.sub.2 R.sup.2, CONHR.sup.12, NHCOR.sup.12, NHCO.sub.2 R.sup.12, NHSO.sub.2 R.sup.12, amidine or guanidine group; PA0 R.sup.5 is H; PA0 R.sup.4 is H, C.sub.1-6 alkyl, aryl, heteroaryl, C.sub.1-2 alkyl-aryl or C.sub.1-2 alkyl-heteroaryl; NR.sup.4 R.sup.5 may form a 5-7 membered ring such as a pyrrolidine, piperidine or morpholine; PA0 R.sup.7 is aryl (optionally substituted with R.sup.10), heteroaryl (optionally substituted with R.sup.10), C.sub.1-4 alkyl-aryl (optionally substituted with R.sup.10), C.sub.1-4 alkyl-heteroaryl (optionally substituted with R.sup.10), C.sub.1-5 alkyl-R.sup.10, C.sub.1-5, alkenyl-R.sup.10, cyclo(C.sub.3-6)alkyl-R.sup.10 ; when Y is C.dbd.O, R7 can also be CO.sub.2 R.sup.2 ; PA0 R.sup.8 is hydrogen; PA0 R.sup.10 is COR.sup.13, NR.sup.9 R.sup.11, N(R.sup.9).sub.2, phthalimido or succinimido; PA0 R.sup.11 is COR.sup.9, CO.sub.2 R.sup.9 (provided R.sup.9 is not H), or SO.sub.2 R.sup.9 (provided R.sup.9 is not H); PA0 R.sup.12 is C.sub.1-4 alkyl, C.sub.1-4 alkylaryl, C.sub.1-4 alkyl-heteroaryl; PA0 and R.sup.13 is OH, OC.sub.1-4 alkyl or N(R.sup.9).sub.2 ; PA0 reaction of an acid chloride of formula (II) ##STR4## Wherein Y is C.dbd.O or SO.sub.2 and R.sup.7 is as defined above, with an amine of formula (III) ##STR5##
Thus, overall the excessive production of soluble CD23 has been implicated in the overproduction of IgE, the hallmark of allergic diseases such as extrinsic asthma, rhinitis, allergic conjunctivitis, eczema, atopic dermatitis and anaphylaxis (Sutton et al, Nature, 366, 1993, 421). Elevated levels of soluble CD23 have also been observed in the serum of patients with chronic B lymphocytic leukaemia (Safarti et al, Blood, 71, 1988, 94), and in the synovial fluid of patients with rheumatoid arthritis (Chomarat et al, Arthritis and Rheumatism, 36, 1993, 234).
It is therefore, a further objective of the present invention to provide compounds which inhibit the formation of human soluble CD23 for the production of a medicament for the treatment or prophylaxis of disorders such as allergy and autoimmune disease in which the overproduction of soluble CD23 is implicated, such as those described above.
Recent reports suggest that new enzymes of the MMP family also mediate the shedding of adhesion molecules such as the selecting, such as L-selectin. These soluble adhesion molecules are implicated in a number of diseases including cancer, autoimmunity and in the inflammatory response. It has been proposed that once cleaved, the selectin bind to particular ligands and this accounts for their biological activity. Thus, drugs that interfere with or prevent binding of the ligands to the selectins will be useful medicaments for treating a variety of the diseases described above. Therefore, it is a yet further objective of the present invention to provide compounds which inhibit the shedding of certain adhesion molecules and thus provide the production of a medicament for the treatment or prophylaxis of disorders such as cancer, autoimmune diseases or inflammatory diseases (such as inflammatory bowel disease and multiple sclerosis).
It is also believed that specific inhibitors of stromelysin and collagenase should be useful as birth control agents. There is evidence that expression of metalloproteinases, including stromelysin and collagenase, is observed in unfertilised eggs and zygotes and at further cleavage stages and increased at the blastocyst stage of fetal development and with endoderm differentiation (C. A. Brenner et al (1989), Genes & Develop., 3:848-59). By analogy to tumour invasion, a blastocyst may express metalloproteinases in order to penetrate the extracellular matrix of the uterine wall during implantation. Inhibition of stromelysin and collagenase during these early development processes should presumably prevent normal embryonic development and/or implantation in the uterus. Such intervention would constitute a novel method of birth control. In addition there is evidence that collagenase is important in ovulation processes. In this example, a covering of collagen over the apical region of the follicle must be penetrated in order for the ovum to escape. Collagenase has been detected during this process and an inhibitor has been shown to be effective in preventing ovulation (J. F. Woessner et al (1989), Steroids, 54:491-499). There may also be a role for stromelysin activity during ovulation (C. K. L. Too et al (1984), Endocrin., 115:1043-1050).
Collagenolytic and stromelysin activity have also been observed in dystrophic epidermolysis bullosa (A. Kronberger et al (1982), J. Invest. Dermatol., 79:208-211; D. Sawamura et al (1991), Biochem. Biophys. Res. Commun., 184:1003-8). Inhibition of metalloendoproteinases should limit the rapid destruction of connective components of the skin.
In addition to extracellular matrix comprising structural components, stromelysin can degrade other in vivo substrates including the inhibitors a.sub.1 -proteinase inhibitor and may therefore influence the activities of other proteinases such as elastase (P. G. Winyard et al (1991), FEBS Letts., 279,1:91-94). Inhibition of the matrix metalloendoproteinases may potentiate the antiproteinase activity of these endogenous inhibitors.
From recent publications it is evident that several new enzymes of the MMP family have been identified, some of which maybe important in disease. Collagenase 3, an enzyme found in to breast carcinoma tissue and other disease states such as arthritis, may have utility in breast cancer (JMP Freije et al (1994), J. Biol. Chem., 269 (24):16766-16773), whilst MT-MMPs, other members of the MMP family have been shown to be key enzymes in the activation of gelatinase A (H Sato et al (1994), Nature, 370:61-65). Gelatinase A is an important enzyme in the growth and metastasis of tumours (such as defined above).
The degradation of .beta.-Amyloid Precusor Protein (APP) has been shown to generate amyloid plaques, a major constituent of the senile plaques, found in patients with Alzheimers Disease (AD). Two recent publications have identified metalloproteinase enzymes that cleave APP to the amyloid plaque (CR Abraham et al (1994), Biochemistry, 33:192-199; G Huber et al (1994), Biochem. Biophys. Res. Comm., 201 (1):45-53).
As appreciated by those of skill in the art, the significant proportion of homology between these new enzymes and other MMPs leads to the possibility that a compound that inhibits one enzyme may to some degree inhibit these new enzymes. Therefore, inhibitors encompassed in this invention may be useful in the diseases in which these new enzymes are implicated.